The present invention discloses a single-tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for early and rapid detection of Zika virus (ZIKV) infection. The single-tube RT-LAMP assay disclosed according to the present invention comprises of a combination of 6 primer sequences or 6 nucleotide fragments represented by sequences (SEQ ID NO: 1) to (SEQ ID NO: 6) for isothermal detection of ZIKV RNA. In another embodiment, the single-tube RTLAMP assay developed in the present invention is specific and sensitive for broad coverage detection of both the Asian and African ZIKV strains. The RT-LAMP assay can greatly enhance diagnosis of Zika in rural clinics and healthcare facilities where resources are limited.